Generation of mtDNA-exchanged cybrids for determination of the effects of mtDNA mutations on tumor phenotypes.

It has been proposed that mutations of mitochondrial DNA (mtDNA) and resultant mitochondrial dysfunction induce various phenotypes, such as mitochondrial diseases, aging, and tumorigenesis. However, it is difficult to conclude whether mtDNA mutations are truly responsible for these phenotypes due to the regulation of the mitochondrial functions by both mtDNA and nuclear DNA. The mtDNA-exchange techniques are very effective to exclude the influence of nuclear DNA mutations on expression of these phenotypes. Using these techniques, we recently showed that specific mtDNA mutations can regulate tumor cell metastasis. In this chapter, we describe the methods to establish the mtDNA-exchanged cell lines (cybrids). Applying this technique will reveal how mtDNA mutations are related to various biological phenomena.